Catalog # S9144 was discontinued on December 15, 2022
The SNAP-Capture Pull Down Resin is used to selectively capture and immobilize a SNAP-tag® fusion protein from solution using agarose beads. The SNAP-Capture Pull Down Resin is prepared by the coupling of SNAP-tag substrate benzylguanine with a highly cross-linked agarose (4%) with excellent physical characteristics.
These beads have a high loading capacity for SNAP-tag fusion proteins
They show very low non-specific absorption of proteins from a complex lysate
They are especially suitable for pull down applications
Two ml of SNAP-Capture Pull Down Resin is sufficient to perform 25 pull down assays using 80 µl of resin suspension per assay.
The SNAP-tag protein labeling system enables
the specific, covalent attachment of virtually any
molecule to a protein of interest. The SNAP-tag
is based on human O6-alkylguanine-DNA-alkyltransferase
(hAGT). SNAP-tag substrates are
fluorophores, biotin or beads conjugated to guanine
or chloropyrimidine leaving groups via a benzyl
linker. In the labeling reaction, the substituted benzyl
group of the substrate is covalently attached to the
SNAP-tag.
There are two steps to the use of this system: sub cloning and expression of the protein of interest as a SNAP-tag fusion, and capture and immobilization of the fusion protein using SNAP-Capture Pull Down Resin.
For larger volume requirements, customized and bulk packaging is available by purchasing through the OEM/Bulks department at NEB. Please contact custom@neb.com for further information.
Figure 1. Substrate structure on SNAP-Capture Pull Down Resin
This product is related to the following categories:
Protein sample containing the protein to immobilize expressed as a SNAP-tag fusion
Buffer for immobilization and washing
Microcentrifuge
Binding Capacity
SNAP-Capture Pull Down Resin (80 µl) was washed, incubated with 200 µl of 1 mg/ml SNAP-tag CBD (Chitin Binding Domain) for 1 hour at room temperature, then rewashed as described in these instructions. Binding capacity was determined to be 1 mg/ml bed resin.
Storage of Pull Down Resin: Do notfreeze the SNAP-Capture Pull Down Resin. Store the unused resin at 4°C. Withproper storage, resin should be stable for at least Two years.
Correct storage and handling of SNAP-tag fusion proteins is essential tomaintain reactivity of the SNAP-tag prior to immobilization.
SNAP-tag fusion proteins can be purified before immobilization, but theimmobilization reaction also works in non-purified protein solutions includingcell lysates.
Add 1 mM DTT to buffers used for the storage of SNAP-tag fusion proteins.Protein samples should be stored at -20°C, or at -80°C for long-term storage.Handling at temperatures above 0°C should be minimized by thawing the SNAP-tagprotein samples shortly before use, and keeping them on ice until just beforethe immobilization.
If a particular fusion protein requires buffers without reducing agents, payparticular attention to minimize all handling steps of the protein above 4°Cbefore the labeling reaction.
The SNAP-tag itself is tolerant of a wide range of buffers. The requirements ofyour fusion partner should dictate the selection of the buffer. The followingstorage buffer composition is recommended, especially when freezing proteinmaterial: pH between 7.0 and 8.0, monovalent salts (e.g. sodium chloride)between 50 mM and 250 mM, and at least 1 mM DTT. Non-ionic detergents can beadded if required, but ionic detergents should be avoided because they reducethe activity of the SNAP-tag. Many proteins benefit from the addition ofglycerol for frozen storage, typically 20% v/v.
Immobilized SNAP-tag fusion proteins should be stored at 4°C. Sodium azide maybe added to 2 mM final concentration to prevent bacterial growth. Depending onthe stability of the fusion partner, under these conditions the immobilizedprotein should be stable at 2–6°C for several months. The SNAP-Capture Pull Down Resin should not be frozen.
The SNAP-tag fusion protein is linked to the SNAP-Capture Pull Down Resin by acovalent bond. Therefore the immobilized protein is essentially irreversiblybound to the resin. It is important however to preserve the functional stabilityof the protein fused to the SNAP-tag as much as possible. We recommend handlingthe immobilized fusion protein and storing between use at 4°C, to prepare itjust before use, and to handle it as gently as possible.
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
Licenses
Notice To Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT PURPOSES ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938.
Cellular Imaging and Analysis (i.e., SNAP and CLIP products)
Notice to Buyer/User: The Buyer/User has a non-exclusive license to use this system or any component thereof for RESEARCH AND DEVELOPMENT ONLY. Commercial use of this system or any components thereof requires a license from New England Biolabs, Inc., 240 County Road, Ipswich, MA 01938.
These patents and patent applications are owned by Covalys, or owned by the Ecole Polytechnique Fédérale de Lausanne (EPFL) and exclusively licensed to Covalys and NEB.
The products and/or their use may be covered by one or more of the following patents and patent applications: Methods for Using O6-Alkylguanine-DNA-Alkyltransferases: US 7,939,284; US 8,367,361; EP 1 410 023; EP 2 037 271; EP 1 696 234; EP 2 211 177; JP 4195815; JP 4226053; CN 1295510; CN 1975422; and SG 100125.
Substrates for O6-Alkylguanine-DNA Alkyltransferase: US 7,799,524; and JP 4976651.
Mutants of O6-Alkylguanine-DNA-Alkyltransferases: US 7,888,090. EP 1720981 (pending).
Specific Substrates for O6-Alkylguanine-DNA-Alkyltransferases: US 8,163,479. EP 1730298 (pending).
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